Is DRAQ7 fixable?
Unlike other commonly used nuclear viability stains such as propidium iodide or DRAQ7™, NucFix™ labeling is extremely stable, allowing the cells to be fixed and permeabilized without loss of fluorescence or dye transfer between cells.
What is DRAQ5?
DRAQ5™ Fluorescent Probe is a far-red DNA stain for fluorescent cellular imaging applications with live or fixed cells. Because of its far-red excitation and emission, the DRAQ5 Stain can be multiplexed with many other fluorophores.
What stains DRAQ7?
DRAQ7 Dye is a membrane-impermeable dye that rapidly stains the double-stranded DNA (dsDNA) of dead or permeabilized cells.
How do I use DRAQ5?
Protocol for DNA staining using DRAQ5™:
- Perform surface staining following protocol of choice.
- Wash cells twice with phosphate buffered saline (PBS).
- Dilute DRAQ5™ to required concentration.
- Incubate at room temperature, preventing exposure to light, for 10-15 minutes.
Is DRAQ5 toxic?
The DRAQ5 is toxic in the sense that it binds DNA and interferes with replication: cells appear to remain viable but be halted in G2/M phase. So if you are after culturing your cells after analysis, then DRAQ5 is not for you.
Does DRAQ5 stain dead cells?
DRAQ5™ is an anthraquinone dye with high affinity for double-stranded DNA. It is a membrane-permeable dye that can label live or fixed/dead cells. In flow cytometry, this dye can be used to distinguish nucleated and non-nucleated cells.
What is phototoxicity in microscopy?
Phototoxicity is the process by which upon illumination, with high laser power or for prolonged periods, the imaged organisms/cells are damaged. Phototoxicity can cause cellular membrane blebbing, vacuole formation and even cell death.
What does DRAQ5 bind?
At higher concentrations (3 and 7.5 microM), DRAQ5 interfered with binding of H2B core histones to DNA.
Can you use DAPI on live cells?
It is used extensively in fluorescence microscopy. As DAPI can pass through an intact cell membrane, it can be used to stain both live and fixed cells, though it passes through the membrane less efficiently in live cells and therefore provides a marker for membrane viability.
How do you test for phototoxicity?
Although specimens show diverse signs of light-induced damage, common themes do exist. A frequent recommendation for assessing phototoxicity is to look for tell- tale morphological signs such as cellular swelling and rounding, blebbing, or the appearance of vacuoles.
How does Hoechst stain work?
Hoechst 33342 binds preferentially to adenine-thymine (A-T) regions of DNA. This stain binds into the minor groove of DNA and exhibits distinct fluorescence emission spectra that are dependent on dye:base pair ratios.
What concentration does DAPI use?
0.1 µg/mL
It is recommended to use DAPI Staining Solution at a final concentration of 0.1 µg/mL. Since application vary, each investigator should titrate the reagent to obtain optimal results.
What is the excitation wavelength for DRAQ7?
DRAQ7™ may be excited by a wide range of wavelengths from 488-647 nm. Despite low absorbance at 488 nm, this excitation source allows for convenient combination with FITC and PE conjugates as well as EGFP. DRAQ7™ is a far-red fluorescent DNA dye that only stains the nuclei in dead and permeabilized cells.
What is DRAQ7 used for?
Find out more. DRAQ7™ is a far-red fluorescent dye that only stains the nuclei in dead and permeabilized cells and can be used in combination with common labels such as GFP or FITC. DRAQ7 is the ideal tool to study dead or membrane-compromised cells because it does not enter intact, live cells.
How do you prepare DRAQ7 for flow cytometry?
Flow Cytometry: Prepare cells for DRAQ7™ staining by resuspending cells in PBS at a concentration of no more than 5 x 10 5 cells/ml. For each 0. 5 ml of cell suspension, dilute DRAQ7™ 1:100 by adding 5 μl to suspended cells, achieving a final concetration of 3 μM. Gently mix and incubate for 10 minutes on ice.